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We narrowed to 132 results for: plasmid for e coli

Showing: 101 - 120 of 132 results

  1. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    Published
    Sept. 26, 2024, 1:15 p.m.
    ...hold the plasmid together, even if there are gaps or flaps remaining. This incomplete plasmid looks like...robust system of homologous recombination found in E. coli, SLIC is a cheap, standardized, and rapid multi-part...cloning uses short sticky ends to insert genes into plasmid backbones, base pairing between these ~4 base overhangs... overhangs isn’t strong enough to hold a plasmid together through transformation, so the cloned vector...longer single-stranded overhangs can hold a nicked plasmid together long enough to get it into cells and let... of LIC is often limited to specially designed plasmids. Sequence- and ligation-independent cloning: A...fixing up the “damage” and producing a complete plasmid. Besides being compatible with any vector, the ...

  2. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    Published
    March 3, 2020, 2:15 p.m.
    ...has been used to construct plasmids for use in model organisms such as E. coli and yeast. Because this approach...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...need. If you'd ever like to write about a recent plasmid deposit please sign up here.        AAV delivery... Figure 1: The base editor is split into two plasmids and co-transfected into cells for reconstitution...overcome this overlap, Yiyang Gong’s lab created two plasmids containing a red-shifted Ace-mScarlet, a fusion...channelrhodopsins. Find the Ace-mScarlet voltage indicator plasmids at Addgene! Beck et al., Sci Report 2019. PubMed...

  3. CRISPR in the Clinic

    Type
    Blog Post
    Published
    Nov. 2, 2023, 1:15 p.m.
    ...infections. The Cas3 enzymes target several strains of E. coli and lethally chop their genomes into many, many...ijms24021563 Resources on Addgene.org CRISPR Plasmids and Resources CRISPR 101 eBook CRISPR History ...Hussein M, Molina MA, Berkhout B, Herrera-Carrillo E. A CRISPR-Cas Cure for HIV/AIDS. Int J Mol Sci. 2023...

  4. Live and Let Dye: Self-Labeling Protein Tags

    Type
    Blog Post
    Published
    April 16, 2024, 1:15 p.m.
    ... demonstrated in 2005 that proteins fused with E. coli dihydrofolate reductase (eDHFR) could be labeled...resources References Bottanelli, F., Kromann, E. B., Allgeyer, E. S., Erdmann, R. S., Wood Baguley, S., Sirinakis...Allgeyer, E. S., Zhong, S., Thompson, A. D., Lowe, N., Butler, R., Bewersdorf, J., Rothman, J. E., Johnston...freezer, you’ll probably find lots of different plasmids containing the same protein of interest tagged...Addgene’s collection, so you may be able to get a plasmid ready-to-use without any cloning at all! As mentioned...Schepartz, A., Baddeley, D., Toomre, D. K., Rothman, J. E., & Bewersdorf, J. (2016). Two-colour live-cell nanoscale....chembiol.2019.01.003 Gallagher, S. S., Sable, J. E., Sheetz, M. P., & Cornish, V. W. (2009). An In Vivo...

  5. A Deep Dive into BioBuilder

    Type
    Blog Post
    Published
    Oct. 11, 2017, 1:44 p.m.
    ...Standards Find Plasmids for Creating and Controlling Gene Networks Browse All SynBio Plasmids ...Standards Synthetic Photobiology: Optogentics for E. coli Browse All Synthetic Biology Blog Posts Resources...

  6. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    Published
    April 11, 2017, 2:30 p.m.
    ...Addgene.org Find the Spinach Plasmids at Addgene Find Broccoli Plasmids at Addgene View All Fluorescent... fluorescence-activated cell sorting (FACS) of E. coli transformed with aptamer libraries, the Jaffrey...PubMed Central PMCID: PMC4800036. 12. Wang, Rongsheng E., et al. “Aptamer-Based Fluorescent Biosensors.” Curr...nucleotide aptamer they called Broccoli. As a product of SELEX and FACS, Broccoli is inherently more stable...new RNA-based scaffolds to contain and stabilize Broccoli cassettes, thereby extending their use into mammalian...mammalian cells. They’ve also dimerized Broccoli to further enhance its biosensor capabilities (see Fig...et al. “In-gel imaging of RNA processing using broccoli reveals optimal aptamer expression strategies....

  7. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    Published
    Jan. 13, 2025, 9:49 p.m.
    ...Duby, J. E., Steinbeck, B. J., McCreary, J., Pierce, S. E., Tolar, J., Meissner, T. B., Chaikof, E. L., Osborn...this evolving field! Find prime editing plasmids! Find plasmids from David Liu's lab! This post was originally...the CRISPR guide Find CRISPR plasmids by function Browse our entire plasmid collection ...prime editors with RT domains derived from the E. coli Ec48 retron RT and the S. pombe Tf1 retrotransposon...for treating genetic diseases. In our summer Hot Plasmids post, we covered the Liu lab’s first demonstration...-01025-z Doman, J. L., Pandey, S., Neugebauer, M. E., An, M., Davis, J. R., Randolph, P. B., McElroy, ...Solley, S. C., Gilbert, L. A., Chan, M. M., Bauer, D. E., Marson, A., Parsons, L. R., & Adamson, B. (2024)...

  8. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

    Type
    Blog Post
    Published
    May 22, 2018, 1:38 p.m.
    ...methionine (i.e. the plasmid complements the deletion). Find the Plasmids Here Plasmid features enable multiple... that allow students to propagate the plasmids in either E. coli (ori and ampR) or S. cerevisiae (2µm ...the overexpression plasmids. Finally, students determine if the presence of the plasmid corrects any known...diverged. Tips on using the Pathways Over Time plasmids The plasmids are used as part of a semester-long research...less-well studied organism into overexpression plasmids. Students next transform deletion strains from...phenotypic abnormalities due to the deletion (i.e. the plasmid complements the deletion). If the phenotype is ...cerevisiae met deletion strains with overexpression plasmids carrying MET/Met genes from either S. pombe or...

  10. Anatomy of a Plasmid Page at Addgene

    Type
    Blog Post
    Published
    Feb. 4, 2016, 3:30 p.m.
    ... best bet is the Plasmid Map: We auto-generate a plasmid map based off of the plasmid sequence and feature...name of the strain.  Most Addgene plasmids are maintained in the E. coli lab strain DH5a, but others may...colleagues. Top of the plasmid page The ‘breadcrumb’ trail at the top of any plasmid page gives the hierarchy... to the individual plasmid page. The depositing lab page contains all the plasmids from the same lab while...contains all the plasmids deposited from the same publication. Next up, we have the plasmid name: This is...other plasmids with similar names in the collection, but you can use this name to find this plasmid from...A quicker way to find this plasmid through search is... The Addgene plasmid number: This is the unique...

  11. Pooled Library Amplifications

    Type
    Blog Post
    Published
    May 30, 2024, 1:30 p.m.
    ...recombined plasmid to overtake full-size plasmids (or for smaller plasmids to overtake larger plasmids, if the...How many unique plasmids does your library contain?), the variance of size of plasmids in your library...transformation amplification protocol is the Chong Zhang E. coli Genome-wide Inhibition Library. DNA and cell amounts...lead to issues such as recombination and loss of plasmid diversity, so Addgene strives to distribute aliquots...library (How important is it that every single plasmid in your library is present, and how will you ensure...30 to 200 times as many colonies as individual plasmids in your library. As an example, if you want 100x... your library, and your library contains 5,000 plasmids, your goal for your transformation would be to...

  12. CRISPR 101: Ribonucleoprotein (RNP) Delivery

    Type
    Blog Post
    Published
    Sept. 6, 2018, 12:02 p.m.
    ...express and purify His-tagged Cas9 from E. coli using a plasmid such as pET-28b-Cas9-His from Alex Schier...commonly performed by transfecting cells with a plasmid, such as PX459, which encodes Cas9 and contains...chosen for both Cas9 and gRNA expression       Plasmid DNA may be incorporated into the genome     Off-target...targets with similar efficiency as compared to plasmid-based expression of Cas9/gRNA and can be used for... as CMV or EF1A promoters found in many CRISPR plasmids) are not expressed. Because this method does ...fluorophore tagged dCas9 may require the use of plasmid or viral-mediated delivery. You’ve decided to...experimental systems that may not be amenable to plasmid based methods. One of the most common techniques...

  13. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    Published
    April 14, 2022, 1:15 p.m.
    ...specificity and binding affinity, can be produced in E. coli, and are both soluble and easily engineered.  Monobodies...Resources Additional resources on the Addgene blog Plasmids 101: Aptamer FluorophoresAntibodies 101: scFVs...available from AddgeneBrowse our recombinant antibody plasmid collection References Grumezescu, A. M. (2018)....

  14. Hot Plasmids - August 2020

    Type
    Blog Post
    Published
    Aug. 18, 2020, 1:15 p.m.
    ...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...They generated plasmids tagged with EB3 to mark the growing tips of microtubuli and plasmids tagged with ...and dCas9-VPR targeting plasmids can be found at Addgene. Check out these plasmids for piggyBac gRNA delivery...highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene, head... Find the plasmids at Addgene The MoClo CRISPR/Cas Toolkit for plants includes 95 plasmids consisting...Nanobodies are easily expressed in the cytosol of E. coli, however the nanobody proteins tend to get stuck...

  15. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    Published
    Oct. 10, 2023, 1 p.m.
    ...resistance DNA plasmids are traditionally grown in bacteria as a way to produce more of the plasmid, isolate...isolate individual plasmids, or express a gene contained in a plasmid. During the propagation and/or transformation...receive a copy of the plasmid or will lose it during division. These non-plasmid-containing bacteria can...Over-Agar Antibiotic Plating Plasmids 101 eBook Resources on the Addgene blog Plasmids 101: Antibiotic Resistance...Resistance Plasmids 101: Blue-White Screening Choosing Your Perfect Plasmid Backbone  ...Plasmids need antibiotic resistance genes so that you can specifically isolate bacteria containing your...your experiment — to grow, express, or isolate plasmids. The presence of an antibiotic resistance gene...

  16. CRISPR Antimicrobials

    Type
    Blog Post
    Published
    May 3, 2016, 2:30 p.m.
    ...resistance genes in E. coli using plasmid and phagemid delivery systems. Phagemids are plasmids containing a ...genome is lethal. If this DSB occurs on a plasmid, the plasmid will be eliminated from the bacterium, which...sequence-specific, antimicrobial, plasmid-based, CRISPR systems (Find the Citorik Plasmids here). In both cases, ...sequence results in cell death. Plasmid targeting may result in plasmid loss and antibiotic resensitization...this resistance gene being encoded by a high-copy plasmid. In the case of the genomically encoded quinolone...RNAs (crRNAs) targeting either genomic (blue) or plasmid-based sequences (red). Treating a mixed population...specifically targets bacteria with the given genomic or plasmid sequence from a mixed population without affecting...

  17. Viral Production at Addgene

    Type
    Blog Post
    Published
    Oct. 24, 2023, 1:15 p.m.
    ...the transfer plasmid, a plasmid encoding rep and serotype-specific cap genes, and a plasmid encoding adenoviral...coli. In addition, plasmids are typically prepared using endotoxin-free plasmid purification kits.   ... all plasmids used for viral production are propagated in the endA-mutated NEB Stable strain of E. coli...contamination is minimized by using an endotoxin-free plasmid purification protocol. To minimize the immunogenic...offline and discarded. Confirmation of the transfer plasmid Addgene uses a rigorous barcode matching system...undergoes PCR with primers targeting the transfer plasmid used in the transfection. PCR products are analyzed... system and PCR-based confirmation of transfer plasmids used in Addgene’s viral production. Barcoding ...

  18. Plasmids 101: Secondary Nanobody Toolbox

    Type
    Blog Post
    Published
    Feb. 27, 2018, 2:04 p.m.
    ...Activity Learn more about protein tags Read about E. coli strains for protein expression Additional Resources... How to Use the Nanobody Toolbox Find Nanobody Plasmids Structural comparison As the name suggests, ... Species Antibody Subclass Nanobody Addgene Plasmid Rabbit N/A* TP897 pTP1183 (1x cysteine) pTP955...Nanobody In Bacteria: Table 2 also has a link to the plasmid you’ll need to express your nanobody of choice ...column. bdNEDP1 protease can be generated using plasmid pDG02583. Label the Nanobodies: Purified nanobodies...Journal of Cell Biology. PubMed PMID: 29263082 Find plasmids from this publication at Addgene. 2. Bates, M... on Addgene.org Find more nanobody expression plasmids here Additional Resources Looking for monoclonal...

  19. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    Published
    April 3, 2014, 8:05 p.m.
    ...trp High levels of gene expression Promoter from E. coli tryptophan operon Repressible Gets turned off with...Once you've gotten to know what a plasmid is in general, it's time to take a look at some of its parts...origin of replication are necessary to replicate plasmids and make sure the cells maintains it. Here, we'll...the promoter, which is necessary for getting the plasmid express our gene of interest. This element is responsible.... This means that the promoter present in your plasmid backbone must to be compatible with the type of...transcript, you will also need to make sure your plasmid has a promoter suited to working in your host organism...development-specific promoters available to scientists. Plasmids are oftentimes put to therapeutic uses, and in...

  20. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    Published
    Oct. 14, 2015, 2:30 p.m.
    ...Francisella Cpf1 (FnCpf1), they employed an E. coli plasmid depletion assay to discover FnCpf1’s PAM sequence...system." Cell (2015). PubMed PMID: 26422227. Find plasmids from this publication at Addgene. 2. Zetsche,....1 (2016): 31-34. PubMed PMID: 27918548. Find plasmids from this publication at Addgene. 3. Kim, Daesik...Biotechnol. (2016). PubMed PMID: 27272384 Find plasmids from this publication at Addgene. Kleinstiver...Biotechnol. (2016). PubMed PMID: 27347757. Find plasmids from this publication at Addgene. Makarova, Kira...PMID: 26411297. Resources at Addgene Find CRISPR plasmids at Addgene Read Addgene's CRISPR Resource Guide...
Showing: 101 - 120 of 132 results